Objective To investigate the expression of microRNA -216a(miR -216a)in patients suf-fering with breast cancer ,and identify the function and mechanism of miR -216 a on autophagy .Methods The expression of miR-216 a in 30 tumor tissues and paired normal tissue of breast cancer were detected by qRT -PCR.Inhibiton of miR-216a in MCF-7 cell lines was done by transfection of miR -216a inhibitor ( AMO-216a).Cell viability was detected by MTT assay .The level of Beclin 1 was detected by western blot .Results The level of miR-216 a was significant elevated in tumor tissue .Cell viability was markedly decreased owing to inhibition of miR-216a in MCF-7 cells.The level of Beclin 1 was significantly increased by transfection of miR-216a inhibitor in MCF-7cells(P<0.05).Conclusion The level of miR-216a is increased in breast canc-er tissue, and it might at least in part promote breast cancer via downregulating Beclin 1 and affecting autophagy .

Objective To explore the relationship among just-world belief,optimism and life satisfaction in colleges.Methods According to convenience sampling principle,370 colleges were selected and assessed with the just world scale for college students (JWS),satisfaction with life scale (SWLS) and the life orientation test-revised (LOT-R) ;and data were analyzed using correlation analysis and regression analysis.Results ①The scores of just-world belief,optimism and life satisfaction were (3.22 ± 0.46),(3.50 ± 0.57),and (3.70 ± 1.21),all of which were higher than middle points.②The scores of just-world belief were positive correlated with the scores of optimism and life satisfaction (r =0.44,0.36,P ＜ 0.01),and the scores of optimism also were positive correlated with life satisfaction(r =0.38,P ＜ 0.01).③ The just-world belief and optimism can positively predict life satisfaction directly(β =0.24,0.29,P ＜ 0.01),and the two factors had interaction(β=0.09,P＜ 0.05).Among optimistic students,life satisfaction of low just-world belief ones were much worse than that of high just world belief ones.However,among pessimistic students,the differences of life satisfaction between high and low just-world belief ones became little.Conclusion Just-world belief of colleges has significant predictive effect on life satisfaction,and the two factors had interaction.

Objective To investigate the expression of peroxisome proliferator-activated receptor-γ(PPAR-γ) and angiotensin receptor (AT2R) in chronic pancreatitis tissue. Methods Between April 2006 and February 2009, 24 patients were pathologically diagnosed as chronic pancreatitis were enrolled and 12 samples of normal pancreatic tissues were treated as controls. Immunohistochemical staining was used to detect PPAR-γ and AT1R, AT2R expression in pancreatic tissues. Results PPAR-γ and AT1R were mostly negatively expressed in normal pancreatic tissues, the positive scores were 0.33±0.49 and 0.42 ± 0. 51, respectively, while AT2R were weakly positively or negatively expressed in acinus and islet cell, and it was weakly positively expressed in ductal epithelial cells, the positive score was 2. 33 ± 1. 37. In chronic pancreatitis tissue, PPAR-γ, AT1R and AT2R were positively expressed in acinus, ductal epithelial cells, and islet cell, the positive scores were 3.28 ± 2.46, 4.36 ± 2.80 and 4.61 ± 2.89, which were significantly higher than those in the normal group (P<0.01, P <0.01, P<0.05). Conclusions PPAR-γ, AT1R and AT2R were positively expressed in chronic pancreatitis tissue, and they may be the treatment target of chronic pancreatitis.

Objective To screen out the polymerase chain reaction ( PCR) primers for specifically identifying Pheretima aspergillum (E. Perrier) , so as to establish a rapid and accurate method to identify the origin of Pheretima aspergillum. Methods Four kinds of earthworms recorded in pharmacopoeia and six kinds of their adulterants commonly seen in the market were collected. The 12SrRNA sequences related to earthworm were downloaded from GenBank database. PCR amplification for ten kinds of samples was performed using the universal primers, and then the products were sequenced. According to the differences between the above sequences, three pairs of specific primers in the non-conservative district were designed for identification of Pheretima aspergillum. Results High-specificity PCR amplification for Pheretima aspergillum with 12St/12Stf primer occurred when the annealing temperature was increased to 64℃, and only Pheretima aspergillum had single amplification strip, while no strips were found in the other adulterants under the same condition. The achieved specific primers could be well verified in 15 kinds of medicinal materials of earthworm purchased in the market, which had the same morphological features showed by the traditional identifying method. Conclusion With 12St/12Stf primer as a specific marker, the identifica tion of Pheretima aspergillum is rapid and accurate in related species of Pheretima aspergillum, and avoids the effect of some factors such as integrity of experimental materials and drying process.

Objective To investigate the medical and pharmaceutical knowledge of patients with chronic diseases and analyse the influence factors of rational administration in patients ,to provid data to support the establishment of pharmaceutical service mode . Methods 386 cases of patients with chronic diseases were asked to finish the questionnaires for the medical and pharmaceutical knowledge ,and factors affecting the rational drug were analyzed by single factor and multiple factors Logistic regression analysis . Results Among the 386 patients ,cardiovascular and celebralvascular disease ratio was the highest(53 .3% ) ,followed by respiratory system diseases(13 .8% ) and the musculoskeletal system diseases (11 .50% );The averaged score of 386 patients was 1 .76 ± 0 .78 , medication knowledge was at a general level;single factor analysis results showed that there was significant difference(P<0 .05) between rational drug-use and abuse of drugs among patients in number ,form of payment ,marital status ,income ,education level , taking drug knowledge lectures ,combined treatment .Multivariate Logistic regression analysis showed that education level ,partici-pation in lectures ,drug combination ,disease species had a significant impact on the rational drug use among patients with chronic disease(P<0 .05) .Conclusion The pharmaceutical knowledge that patients with chronic disease mastered is unsatisfactory ;and unreasonable behavior of medication is common scence .Education level ,participation in lectures ,drug combination ,the number of diseases have great influence on the rational use of drugs in patients with chronic diseases .A kind of effective pharmaceutical service mode should be established for patients with chronic diseases by clinical pharmacists .This is a very meaningful work for rational ad-ministration .

Objective To study the transfection effects of nuclear factor-KappaB(NF-κB)decoy oligodeoxynucleotides(ODN) to Kupffer cells (KCs) mediated by lipofectamine,and investigate it's suppression effects on KCs activation. Methods Twenty-four Wistar rats were divided into three groups (n=8).(1)Control group,in which the normal KCs were isolated.(2)LPS group,in which 1 ms/L LPs was added to the culture system.(3)NF-κB decoy ODN group,in which KCs were transduced with NF-κB decoy ODN (4μg×105KCs)prior to LPS stimulation.The transfection efficiency Was assayed,and the phagocytosis function,NF-κB(P65) translocation,CD40 mRNA expression of KCs were also detected respectively. Results Kupffer cells were obviously activated after LPS stimulation.the phagocytosis function was reinforced.the activity of NF-κB transloeated from cytoplasm into nucleus was obviosly increaced.The co-stimulatory molecules expression(CD40 mRNA)significantly increased compared with control group(t=4.01,P<0.01).NF-κB decoy oligodeoxynucleotides can efficiently transfected into KCs mediated by lipofectamine,which can obviously suppress KCs activation,and downregulate the expression of downstream gene(compared with LPS group,t=4.89,P<0.01). Condusion NF-κB decoy ODN can efficiently transfect into KCs and inhibit it's activation.

Lanosterol synthase is encoded by the erg7 gene and catalyzes the cyclization of 2, 3-oxidosqualene, which is a rate-limiting step of the inherent mevalonate (MVA)/ergosterol metabolic pathway in Saccharomyces cerevisiae. The intermediate 2, 3-oxidosqualene is also the precursor of triterpenoids. Therefore, the cyclization of 2, 3-oxidosqualene is the key branch point of ergosterol and triterpenoids biosynthesis. Down-regulation of 2, 3-oxidosqualene metabolic flux to ergosterol in S. cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway reconstructed by the synthetic biology approach. To construct erg7 knockout cassette harboring the loxP-Marker-loxP element, long primers were designed, which were homologous to the sequences of both erg7 ORF and plasmid pUG66. The cassette was transformed into diploid wild strain INVSc1 by LiAc/SS Carrier DNA/PEG method and then erg7 gene haploid deficient mutant was obtained by homologous recombination. The results of semiquantitative PCR and real-time quantitative PCR revealed that erg7 expression level in erg7 gene haploid deficient mutant is one time lower than that in wild strain. The results of TLC and HPLC showed that the ergosterol content in deficient mutant decreased to 42% of that in wild strain.
Chromatography, High Pressure Liquid ; DNA Primers ; Down-Regulation ; Ergosterol ; metabolism ; Haploidy ; Intramolecular Transferases ; genetics ; Polymerase Chain Reaction ; Saccharomyces cerevisiae ; genetics ; Squalene ; analogs & derivatives ; metabolism

Pichia pastoris is one of the most important systems used in the field of molecular biology for the expression of recombinant proteins. The system has advantages of high expression, high stability, high secretion, easy high-density fermentation and low cost. Many factors affect the expression of recombinant protein, such as gene copy number, codon usage preference, type of promoter, molecular chaperones, glycosylation, signal peptide and fermentation process. In this review, research advances of the above aspects are summarized, which lay a foundation for improving the expression of recombinant proteins in P. pastoris.
Fermentation ; Gene Dosage ; Glycosylation ; Molecular Chaperones ; Pichia ; metabolism ; Promoter Regions, Genetic ; Protein Sorting Signals ; Recombinant Proteins ; biosynthesis

Objectives To evaluate the effect of thyroid hormone therapy with low dose of thyroxin on cardiac function in elderly patients with heart failure and sick euthyroid syndrome. Methods Forty-seven patients (33 males and 14 females, mean age 85.9+4.6 years,ranging from 80 to 99 years) with chronic heart failure (NYHA Ⅱ-Ⅳ) and low triiodothyronine (T3) state were randomly allocated to the treatment group or control group. The treatment group patients received oral administration of levothyroxine sodium (Euthyrox) 25-50mg/d in addition to conventional therapy of heart failure, whereas patients in control group were given conventional therapy only. Serum level of total T3 (TT3), free T3 (FT3), total thyroxine (TT4), free thyroxine (FT4), and thyroid-stimulating hormone (TSH) were determined. For both groups, left ventricular ejection fraction (LVEF) and stroke volume (SV) were assessed by two-dimensional echocardiography before and at 8 weeks after treatment. The changes of these parameters after the treatment were evaluated by adjusting heart rate in the two groups. Results The reduced serum T3 level in the treatment group was corrected after thyroid hormone therapy,and these patients had a significant improvement in cardiac function after treatment. By contrast, in the control group only changes of serum TT3 and TT4 levels and SV and LVEF after treatment were statistically significant. The heart rate-adjusted mean SV and LVEF in both groups were also increased, which was significantly greater in the treatment group than in the control group. Conclusion In the elderly patients with heart failure and sick euthyroid syndrome, addition of thyroxine at a low dosage to the conventional treatment could effectively improve the low T3 state and cardiac function independent of changes of heart rate.

Objective To study the activation of alveolar macrophage β (AM) and the expression of co-stimulatory molecule CD40 in transfusion-related acute lung injury (TRALI) model in order to illustrate the pathogenesis of TRALI.Methods Sixty SD rats were randomly (random number) divided into normal control group (n =15) with sham operation using normal saline instead of LPS and plasma,positive control group (n =15) with ALI induced by intravenous infusion of 5 mg/kg lipopolysaccharide (LPS) in equivalent volume of whole blood drawn out),and TRALI group (n =15) treated by intra-peritoneal injection of 2 mg/kg LPS 2 h before the transfusion of human plasma (1 mL whole blood about 10％ of total blood volume drawn out and replaced with 1 mL plasma),LPS control group (n =15) treated by intra-peritoneal injection of 2 mg/kg LPS 2 h before saline infusion in equivalent volume of blood drawn out.The pathologic changes of rat lung tissue were observed by HE staining.The expression of TLR4 was examined by RT-PCR.The activation of NF-κB in AM was measured by electrophoresis mobility shift assay (EMSA).The expression of CD40 mRNA and CD40 molecule were analyzed by Northern blot and flow cytometry respectively.ELISA was performed to detect the concentration of TNF-α,MIP-2 and IL-1 β in broncho-alveolar lavage fluid (BALF).Results Broken alveolar septa,hyperemia,and massive infiltration of inflammatory cells including the neutrophils were observed in lung tissues of TRALI group.The expression of TLR4 gene was detected in activated macrophage phi (AMφ) of TRALI group rats.The activation of NF-κB was increased in TRALI group rats.The expression of CD40 in AMφ was higher in rats of TRALI group than that in rats of control group and LPS control group.The concentration of TNF-αt,MIP-2 and IL-1β were enhanced significantly in BALF of TRALI group rats.Conclusion The activation of AM and up-regulation of costimulatory molecule CD40 induced release of some inflammatory cytokines.It suggested that AM activation may play an important role in the pathogenesis of TRALI.